Infantile outcome at 3 years of age among monochorionic twins with Type-II or -III selective fetal growth restriction with isolated oligohydramnios who underwent fetoscopic laser photocoagulation.

OBJECTIVE: This study aimed to examine infantile outcomes at 3 years of age with selective fetal growth restriction (sFGR) Types II and III with isolated oligohydramnios who underwent fetoscopic laser photocoagulation (FLP). METHODS: This multicenter prospective cohort study included monochorionic diamniotic twins who underwent FLP for sFGR between 16 and 25 weeks of gestation. The indication for performing FLP was in cases of sFGR Type II or III with oligohydramnios, where the maximal vertical pocket was ≤2 cm among twins with FGR. This was done in the absence of a typical twin-twin transfusion syndrome diagnosis. The primary outcome was the intact survival (IS) rate of infants at the corrected age of 40 weeks and 3 years. IS at the corrected age of 40 weeks was defined as survival without grade III or IV intraventricular hemorrhage or cystic periventricular leukomalacia, and IS at 3 years of age was defined as survival without neurodevelopmental morbidity, including cerebral palsy, neurodevelopmental impairment with a total developmental quotient of ≤70, bilateral deafness, or bilateral blindness. RESULTS: Among 45 patients with sFGR, 30 (66.7%) were classified as having Type II and 15 (33.3%) as Type III sFGR. The prevalence of IS at the corrected age of 40 weeks was 51.1% (n=23) in FGR twins and 95.5% (n=42) in larger twins. The prevalence of IS at 3 years of age was 46.7% (n=21) in FGR twins and 86.4% (n=38) in larger twins. Among the 24 FGR twins who were not diagnosed with IS at 3 years of age, 91.7% (22 of 24 cases) suffered fetal or infantile demise other than miscarriage and neurodevelopmental impairment. All larger twins who were not diagnosed with IS at 3 years of age (n=6, 13.6%) had neurological morbidity, in addition to one case of miscarriage. CONCLUSIONS: FGR twins and larger twins, when subjected to FLP due to sFGR coupled with umbilical artery Doppler abnormalities and isolated oligohydramnios, exhibit low rates of neurological morbidity and low mortality, respectively. Therefore, FLP for Type II or III sFGR with oligohydramnios may be a feasible and preferable management option. This article is protected by copyright. All rights reserved.

Combination of fluorescence visualization and iodine solution-guided surgery for local control of early tongue cancer.

The control of enclosed oral epithelial dysplasia is important for the control of oral cancer. Fluorescence visualization and iodine solution are able to detect oral epithelial dysplasia and surrounding oral cancer. The purpose of this study was to clarify the effectiveness of combining fluorescence visualization and iodine solution-guided surgery for early tongue cancer. Participants comprised 264 patients with primary early tongue cancer who underwent surgery. The surgical margin was set at 10 mm outside the clinical tumour, and 5 mm outside the area of fluorescence visualization loss, and 5mm outside the iodine unstained area. The 5-year disease-free survival rate was 87.1% vs 76.1% (P = 0.016) and the 5-year local control rate was 98.6% vs 93.0% (P = 0.008) for combination-guided surgery when compared to conventional surgery. Positive margin rates were 0% for cancer, and 6.5% and 0% for low- and high-grade dysplasia, respectively, with combination-guided surgery (P = 0.257). Multivariate analysis revealed that combination-guided surgery (odds ratio 0.140, 95% confidence interval 0.045-0.437; P < 0.001) and intraoperative frozen section examination (odds ratio 0.302; 95% confidence interval 0.115-0.791; P = 0.015) were significantly associated with local control. The combination of fluorescence visualization and iodine solution are effective in selecting surgical margins for early tongue cancer.

Development of a targeted gene panel for the diagnosis of Gorlin syndrome.

Gorlin syndrome is a rare autosomal dominant disease caused by mutations in the PTCH1, PTCH2, and SUFU genes. Each symptom of the disease has a different time point of onset, which makes early diagnosis based solely on symptoms challenging. In this study, a gene panel was developed to overcome the challenges in the diagnosis of Gorlin syndrome and allow diagnosis using a single test. A custom panel was generated for four genes associated with Gorlin syndrome: PTCH1, PTCH2, SMO, and SUFU. Twenty-seven samples from 12 patients with Gorlin syndrome and three asymptomatic blood relatives of the patients were examined. This panel was highly reliable with a high Q30 quality score, on-target ratio, and coverage. The panel was time- and cost-efficient and enabled the detection of more mutations than whole-exome sequencing for the same patient. Pathogenic mutations in both PTCH1 and PTCH2 were detected in five of the 12 patients with Gorlin syndrome who were diagnosed based on clinical symptoms. Using this panel, the same mutation was identified in the patients and their blood relatives. In summary, this panel facilitated the highly reliable genetic diagnosis of Gorlin syndrome at a low cost, using only blood samples.

An outbreak of food poisoning due to Escherichia coli serotype O7:H4 carrying astA for enteroaggregative E. coli heat-stable enterotoxin1 (EAST1).

In June 2020, a large-scale food poisoning outbreak involving about 3000 elementary and junior high school students occurred in Yashio, Saitama, Japan. A school lunch was the only food stuff ingested by all of the patients. Escherichia coli serotype O7:H4 carrying the astA gene for enteroaggregative E. coli (EAggEC) heat-stable enterotoxin 1 (EAST1) was detected in faecal specimens from the patients, and sample inspection revealed its presence in a seaweed salad and red seaweed (Gigartina tenella) as one of the raw materials. Analysis of the antibiotic sensitivity of the isolates revealed resistance to ampicillin and cefotaxime. All isolates were confirmed to be of the same origin by pulsed-field gel electrophoresis after digestion with the restriction enzyme XbaI, and single nucleotide polymorphism analysis using whole genome sequencing. To our knowledge, this is the first report of a large-scale food poisoning caused by E. coli O7:H4, which lacks well-characterized virulence genes other than astA.

Functional expression of equilibrative and concentrative nucleoside transporters in alveolar epithelial cells.

Equilibrative nucleoside transporters (ENTs) and concentrative nucleoside transporters (CNTs) mediate the cellular uptake of nucleosides and nucleobases across the plasma membrane and play important roles in the salvage pathways of nucleotide synthesis. However, information about nucleoside transport systems in the lung alveolar epithelial cells is limited. Therefore, in the present study, we examined the function and expression of nucleoside transporters using primary cultured alveolar type II cells and transdifferentiated type I-like cells. The uptake of uridine, a substrate for ENTs and CNTs, in type II and type I-like cells was time, temperature, and concentration dependent, and was inhibited by other nucleoside transporter substrates such as adenosine. Uridine uptake in both cells was insensitive to nanomolar concentrations of NBMPR, a potent ENT1 inhibitor, while it was inhibited by higher concentrations of NBMPR, suggesting that ENT2, but not ENT1, is involved in uridine uptake in these cells. Additionally, uridine uptake was higher in the presence of Na+ than in the absence of Na + and was partially inhibited by a CNT inhibitor phloridzin in these cells, suggesting that CNT is also involved in uridine uptake. In both cells, the mRNA expression of ENT1, ENT2, CNT2, and CNT3 was observed. Finally, the activity of uridine uptake was considerably higher in type II cells than in type I-like cells. In addition, the mRNA expression of ENT2, CNT2, and CNT3, but not ENT1, was lower in type I-like cells than in type II cells. These findings would help understand the functional roles of equilibrative and concentrative nucleoside transporters in alveolar epithelial cells.

Transport of ribavirin in human myelogenous leukemia cell line K562.

The anticancer effect of ribavirin, a purine nucleoside analogue, has been studied using cultured cancer cells such as the human myelogenous leukemia cell line K562. In order to exert its pharmacological effect, ribavirin has to enter cancer cells. However, there is little information concerning the transport mechanism of ribavirin into K562 cells. In this study, therefore, we examined the uptake mechanism of ribavirin in K562 cells. The uptake of ribavirin in K562 cells was time- and temperature-dependent, and was saturable with a Km value of 1.5 mM. Ribavirin uptake was inhibited by nucleosides such as adenosine and uridine, and by inhibitors of equilibrative nucleoside transporter 1 (ENT1) such as S-(4-nitrobenzyl)-6-thioinosine and dipyridamole in a concentration-dependent manner. In addition, the expression of ENT1 mRNA in K562 cells was confirmed by real-time PCR. On the other hand, Na+-dependence of ribavirin uptake was not observed, suggesting the involvement of ENT1, but not Na+-dependent concentrative nucleoside transporters, in ribavirin uptake in K562 cells. Treatment of K562 cells with sodium butyrate induced erythroid differentiation, but ribavirin uptake activity and sensitivity of the uptake to various inhibitors were not different between native and differentiated K562 cells. These results suggest that ribavirin uptake into K562 cells is mainly mediated by ENT1, which may have a pivotal role in anticancer effect of ribavirin.

Functional expression of breast cancer resistance protein and cholesterol effect in human erythrocyte membranes.

In human erythrocyte membranes, various influx and efflux transporters are functionally expressed. However, their transport characteristics and modulation under disease states are not fully understood. In this study, we first examined the expression and detailed transport characteristics of breast cancer resistance protein (BCRP), an efflux ABC transporter, using inside-out membrane vesicles (IOVs) prepared from human erythrocytes, and then studied the effect of membrane cholesterol on BCRP function. The expression of BCRP was confirmed by western blotting; most of them being homodimers. The uptake of lucifer yellow (LY), a fluorescent BCRP substrate, into IOVs was time-, temperature-, and ATP-dependent, and the concentration of ATP which induced half-maximal stimulation of LY uptake was calculated to be 0.39 mM. The uptake of LY by IOVs was saturable with a Km value of 166 µM, and was inhibited by various BCRP inhibitors and substrates, such as fumitremorgin C and mitoxantrone. When membrane cholesterol content was increased by treating IOVs with cholesteryl hemisuccinate, LY uptake decreased with increasing cholesterol content. These results suggest that transport activity of BCRP in human erythrocyte membranes may be suppressed under disease states, such as hypercholesterolemia, that increase membrane cholesterol content.

Reappraisal of classification of distal cholangiocarcinoma based on tumour depth.

BACKGROUND: In the eighth edition of the AJCC cancer staging classification, the T system for distal cholangiocarcinoma (DCC) has been revised from a layer-based to a depth-based approach. The aim of this study was to propose an optimal T classification using a measured depth in resectable DCC. METHODS: Patients who underwent pancreatoduodenectomy for DCC at 32 hospitals between 2001 and 2010 were included. The distance between the level of the naive bile duct and the deepest cancer cells was measured as depth of invasion (DOI). Invasive cancer foci were measured as invasive tumour thickness (ITT). Log rank χ2 scores were used to determine the cut-off points, and concordance index (C-index) to assess the survival discrimination of each T system. RESULTS: Among 404 patients, DOI was measurable in 182 (45·0 per cent) and ITT was measurable in all patients, with median values of 2·3 and 5·6 mm respectively. ITT showed a positive correlation with DOI (rS = 0·854, P < 0·001), and the cut-off points for prognosis were 1, 5 and 10 mm. Median survival time was shorter with increased ITT: 12·4 years for ITT below 1 mm, 5·2 years for ITT at least 1 mm but less than 5 mm, 3·0 years for ITT at least 5 mm but less than 10 mm, and 1·5 years for ITT 10 mm or more (P < 0·001). This classification exhibited more favourable prognostic discrimination than the T systems of the seventh and eighth editions of the AJCC (C-index 0·646, 0·622 and 0·624 respectively). CONCLUSION: ITT is an accurate approach for depth assessment in DCC. The four-tier ITT classification with cut-off points of 1, 5 and 10 mm seems to be a better T system than those in the seventh and eighth editions of the AJCC classification.

P-gp modulating effect of Azadirachta indica extract in multidrug-resistant cancer cell lines.

The extract of Azadirachta indica, commonly known as neem, has found extensive use in traditional medicine for treating various human diseases. In this study, the effect of the 50% ethanol extract of A. indica (AI01) on P-glycoprotein (P-gp)-mediated multidrug resistance (MDR) was examined using MDR cell lines, specifically paclitaxel-resistant HepG2 (PR-HepG2) and doxorubicin (DOX)-resistant (DR) colon-26 cells. 96-h treatment of the two cell lines with AI01 (30 µg/mL) showed no effect on the expression of P-gp mRNA (human MDR1 and mouse mdr1b) and protein, while AI01 increased the accumulation of rhodamine 123, a P-gp substrate, in both PR-HepG2 and DR-colon-26 cells. The cytotoxic effects of 48-h treatment with AI01 on the viability of PR-HepG2 and DR-colon-26 cells were not observed. Therefore, 30 µg/mL AI01 may have no cytotoxic and P-gp-inducing effects. Finally, AI01 potentiated the sensitivity of PR-HepG2 and DR-colon-26 cell lines to DOX by 8.6- and 15.3-fold, respectively. These findings suggest that A. indica may be a promising source for a new class of P-gp modulators without cytotoxic/P-gp induction effects.

Assessment of diastolic function of normal fetal heart using dual-gate Doppler.

OBJECTIVES: The ratio of mitral peak early diastolic filling to early diastolic mitral annular velocity (E/e') reflects diastolic cardiac function in adults. Dual-gate Doppler (DD) enables measurements of E/e' in the same heart beat. This study was designed to assess the utility of the DD method for measurement of fetal E/e' and determine reference ranges for normal fetuses. METHODS: This prospective study comprised normal singleton pregnancies undergoing fetal echocardiography between 16 and 36 weeks of gestation. According to the DD method, E-wave velocity on pulsed-wave Doppler and e'-wave on tissue Doppler imaging were measured simultaneously on an apical or basal four-chamber view, and fetal E/e' was calculated. Spearman's correlation coefficient was used to assess the relationship between gestational age (GA) and E-wave and e'-wave velocities and E/e'. RESULTS: A total of 133 pregnancies were included in this study and all E/e' measurements were successful. Significant correlation was observed between GA and both left ventricular (LV) E/e' (r s  = -0.666, P < 0.001) and right ventricular (RV) E/e' (r s  = -0.607, P < 0.001). The regression equations for bilateral E/e' were: LV-E/e' = 17.341 - 0.631GA + 0.008 × GA2 (mean ± SD, R 2  = 0.440 ± 1.333); and RV-E/e' = 19.156 - 0.794GA + 0.012GA2 (R 2  = 0.419 ± 1.329). CONCLUSIONS: Bilateral E/e' of normal fetuses, measured using the DD method, decreased with GA, which is considered to be related to myocardial maturity. DD is a useful and convenient method for evaluating fetal E/e' in order to assess diastolic function in the prenatal period. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Erratum to: Usefulness and capability of three-dimensional, full high-definition movies for surgical education.

[This corrects the article DOI: 10.1186/s40902-017-0107-3.].

Usefulness and capability of three-dimensional, full high-definition movies for surgical education.

BACKGROUND: Because of changing surgical procedures in the fields of oral and maxillofacial surgery, new methods for surgical education are needed and could include recent advances in digital technology. Many doctors have attempted to use digital technology as educational tools for surgical training, and movies have played an important role in these attempts. We have been using a 3D full high-definition (full-HD) camcorder to record movies of intra-oral surgeries. METHOD: The subjects were medical students and doctors receiving surgical training who did not have actual surgical experience (n = 67). Participants watched an 8-min, 2D movie of orthognathic surgery and subsequently watched the 3D version. After watching the 3D movie, participants were asked to complete a questionnaire. RESULT: A lot of participants (84%) felt a 3D movie excellent or good and answered that the advantages of a 3D movie were their appearance of solidity or realism. Almost all participants (99%) answered that 3D movies were quite useful or useful for medical practice. CONCLUSIONS: Three-dimensional full-HD movies have the potential to improve the quality of medical education and clinical practice in oral and maxillofacial surgery.

Effect of Curcuma comosa extracts on the functions of peptide transporter and P-glycoprotein in intestinal epithelial cells.

Curcuma comosa has been widely used as a herbal medicine in Thailand; however, it remains unclear whether C. comosa influences the absorption of drugs that are substrates for the transporters in the small intestine. In this study, we investigated the effect of C. comosa extracts on the functioning of peptide transporter 1 (PEPT1), an influx transporter, and P-glycoprotein (P-gp), an efflux transporter, in Caco-2 cells and rat intestine. In Caco-2 cells, the ethanolic extract of C. comosa (CCE) lowered the uptake of glycylsarcosine (Gly-Sar), a PEPT1 substrate, while it enhanced the uptake of rhodamine 123 (Rho123), a P-gp substrate, in a concentrationdependent manner. In addition, CCE inhibited apical-to-basal transport of Gly-Sar and basal-to-apical transport of Rho123. Furthermore, the absorption of cephalexin, another PEPT1 substrate, and the exsorption of Rho123 across the rat intestine were inhibited by CCE. Conversely, CCW, the hot water extract of C. comosa, suppresses the function of PEPT1 but not of P-gp in Caco-2 cells. These results suggest that C. comosa used as a herbal medicine in Thailand may affect the intestinal absorption of certain drugs.

Dedifferentiated endometrioid adenocarcinoma of the uterus: a case report.

BACKGROUND: Dedifferentiated endometrioid adenocarcinoma (DEAC) of the uterus was first described by Silva et al. in 2006. The tumor has high-grade endometrial carcinoma component which abruptly emerged from low-grade areas. DEAC showed more aggressive phenotype than FIGO grade 3 endometrioid adenocarcinoma. However, there have been a few studies evaluating effectiveness of adjuvant therapy for the patients with DEC. CASE REPORT: A 41-year-old case with Stage IVB DEAC that clinically showed resistance to several regimens of chemotherapy is reported. The uterine corpus tumor with size of 120 x 100 mm, and the metastases were found in lung, liver, and pelvic lymph nodes. She underwent supra-vaginal hysterectomy, left salpingo-oophorectomy, and partial resection of ileum. Pathologically, the tumor had both well differentiated and undifferentiated carcinoma components, and it was diagnosed as DEAC. After primary surgery, the patient received four regimens of adjuvant chemotherapy, however all regimens were judged as progressive disease. Subsequently, the patient died of disease seven months after surgery. CONCLUSION: The present case of DEAC had an exceedingly poor prognosis, as was suggested in the several previous reports. The review of adjuvant therapeutic modalities revealed that there has been no effective therapy in the response-evaluable patients with DEAC. Further investigations for new strategy to treat the cases with DEAC are needed.

Successful control of an outbreak of GES-5 extended-spectrum ß-lactamase-producing Pseudomonas aeruginosa in a long-term care facility in Japan.

BACKGROUND: Little is known about multidrug-resistant Pseudomonas aeruginosa (MDRP) outbreaks in long-term care facilities (LTCFs). AIM: To describe an MDRP outbreak in an LTCF and to clarify risk factors for MDRP acquisition. METHODS: Patients who were positive for MDRP at an LTCF from January 2013 to January 2014 were analysed. A descriptive analysis, a case-control study, and a microbiological analysis were performed. FINDINGS: A total of 23 MDRP cases were identified, 16 of which were confirmed in sputum samples. Healthcare workers were observed violating hand hygiene procedures when performing oral, wound, and genital care. Nasogastric tube and oxygen mask use was associated with MDRP acquisition in the respiratory tract, which might have been confounded by poor hand hygiene. Sharing unhygienic devices, such as portable oral suction devices for oral care, and washing bottles and ointments for wound and genital care with inadequate disinfection could explain the transmission of MDRP in some cases. Isolates from 11 patients were found to be indistinguishable or closely related by pulsed-field gel electrophoresis and harbouring the blaGES-5 gene. Subsequent enhanced infection control measures were supported by nearby hospitals and a local public health centre. No additional cases were identified for a year after the last case occurred in January 2014. CONCLUSION: An outbreak of MDRP with an antimicrobial resistance gene, blaGES-5, occurred in a Japanese LTCF. It was successfully controlled by enhanced infection control measures, which neighbouring hospitals and a local public health centre supported.

High-temperature-required protein A2 as a predictive marker for response to chemotherapy and prognosis in patients with high-grade serous ovarian cancers.

BACKGROUND: High-temperature-required protein A2 (HtrA2), a protein relating with apoptosis in a caspases-dependent and non-dependent manner, has been reported to be associated with chemosensitivity in several human cancers. METHODS: Tissue microarrays made from 142 patients with high-grade serous ovarian adenocarcinoma were evaluated to assess whether HtrA2 expression was related with several clinical parameters. RESULTS: Negative HtrA2 expression was observed in 36 cases (25%) of the patients, and related with significantly lower response rates of primary chemotherapy than those with positive HtrA2 expression (56% vs 83%, P<0.01). In addition, negative HtrA2 expression was identified as an independent worse prognostic factor for progression-free survival and overall survival by multivariate analyses. Furthermore, HtrA2 downregulation modulated sensitivity to platinum in serous ovarian cancer cells in vitro. CONCLUSIONS: HtrA2 expression was a predictor for sensitivity to chemotherapy, and could be a candidate of molecular target in the treatment of high-grade serous ovarian cancers.